Synthesis of new methoxy derivatives of trans 2,3-diaryl-2,3-dihydrobenzofurans and evaluation of their anti-inflammatory activity.

In the present study we synthesized new methoxy derivatives of trans 2,3-diaryl-2,3-dihydrobenzofurans, starting from suitable trans 2,3-diaryloxiranes, using regio- and stereoselective nucleophilic oxiranyl ring-opening reactions. The compounds were tested as anti-inflammatories in U937 cells. All compounds showed a significant role in inhibiting the NF-κB pathway and were able to restore normal ROS and NO level upon LPS activation. Moreover, regarding inhibition of ACLY, enantioenriched (50% ee) 7a50 showed more potency than the racemic counterpart 7arac, together with a higher reduction of prostaglandin E2 production, thus suggesting a stereoselective interaction in this pathway.

Refinement of the Listeria monocytogenes σB regulon through quantitative proteomic analysis.

σ(B) is an alternative σ factor that regulates stress response and virulence genes in the foodborne pathogen Listeria monocytogenes. To gain further insight into σ(B)-dependent regulatory mechanisms in L. monocytogenes, we (i) performed quantitative proteomic comparisons between the L. monocytogenes parent strain 10403S and an isogenic ΔsigB mutant and (ii) conducted a meta-analysis of published microarray studies on the 10403S σ(B) regulon. A total of 134 genes were found to be significantly positively regulated by σ(B) at the transcriptomic level with >75 % of these genes preceded by putative σ(B)-dependent promoters; 21 of these 134 genes were also found to be positively regulated by σ(B) through proteomics. In addition, 15 proteins were only found to be positively regulated by σ(B) through proteomics analyses, including Lmo1349, a putative glycine cleavage system protein. The lmo1349 gene is preceded by a 5' UTR that functions as a glycine riboswitch, which suggests regulation of glycine metabolism by σ(B) in L. monocytogenes. Herein, we propose a model where σ(B) upregulates pathways that facilitate biosynthesis and uptake of glycine, which may then activate this riboswitch. Our data also (i) identified a number of σ(B)-dependent proteins that appear to be encoded by genes that are co-regulated by multiple transcriptional regulators, in particular PrfA, and (ii) found σ(B)-dependent genes and proteins to be overrepresented in the 'energy metabolism' role category, highlighting contributions of the σ(B) regulon to L. monocytogenes energy metabolism as well as a role of PrfA and σ(B) interaction in regulating aspects of energy metabolism in L. monocytogenes.

How Can AI Help Improve Food Safety?

With advances in artificial intelligence (AI) technologies, the development and implementation of digital food systems are becoming increasingly possible. There is tremendous interest in using different AI applications, such as machine learning models, natural language processing, and computer vision to improve food safety. Possible AI applications are broad and include, but are not limited to, (a) food safety risk prediction and monitoring as well as food safety optimization throughout the supply chain, (b) improved public health systems (e.g., by providing early warning of outbreaks and source attribution), and (c) detection, identification, and characterization of foodborne pathogens. However, AI technologies in food safety lag behind in commercial development because of obstacles such as limited data sharing and limited collaborative research and development efforts. Future actions should be directed toward applying data privacy protection methods, improving data standardization, and developing a collaborative ecosystem to drive innovations in AI applications to food safety.

Phylogeographic Clustering Suggests that Distinct Clades of Salmonella enterica Serovar Mississippi Are Endemic in Australia, the United Kingdom, and the United States.

Salmonella enterica serovar Mississippi is the 2nd and 14th leading cause of human clinical salmonellosis in the Australian island state of Tasmania and the United States, respectively. Despite its public health relevance, relatively little is known about this serovar. Comparison of whole-genome sequence (WGS) data of S. Mississippi isolates with WGS data for 317 additional S. enterica serovars placed one clade of S. Mississippi within S. enterica clade B ("clade B Mississippi") and the other within section Typhi in S. enterica clade A ("clade A Mississippi"), suggesting that these clades evolved from different ancestors. Phylogenetic analysis of 364 S. Mississippi isolates from Australia, the United Kingdom, and the United States suggested that the isolates cluster geographically, with U.S. and Australian isolates representing different subclades (Ai and Aii, respectively) within clade A Mississippi and clade B isolates representing the predominant S. Mississippi isolates in the United Kingdom. Intraclade comparisons suggested that different mobile elements, some of which encode virulence factors, are responsible for the observed differences in gene content among isolates within these clades. Specifically, genetic differences among clade A isolates reflect differences in prophage contents, while differences among clade B isolates are due to the acquisition of a 47.1-kb integrative conjugative element (ICE). Phylogenies inferred from antigenic components (fliC, fljB, and O-antigen-processing genes) support that clade A and B Mississippi isolates acquired these loci from different ancestral serovars. Overall, these data support that different S. Mississippi phylogenetic clades are endemic in Australia, the United Kingdom, and the United States. IMPORTANCE The number of known so-called "polyphyletic" serovars (i.e., phylogenetically distinct clades with the same O and H antigenic formulas) continues to increase as additional Salmonella isolates are sequenced. While serotyping remains a valuable tool for reporting and monitoring Salmonella, more discriminatory analyses for classifying polyphyletic serovars may improve surveillance efforts for these serovars, as we found that for S. Mississippi, distinct genotypes predominate at different geographic locations. Our results suggest that the acquisition of genes encoding O and H antigens from different ancestors led to the emergence of two Mississippi clades. Furthermore, our results suggest that different mobile elements contribute to the microevolution and diversification of isolates within these two clades, which has implications for the acquisition of novel adaptations, such as virulence factors.

Listeria monocytogenes {sigma}B has a small core regulon and a conserved role in virulence but makes differential contributions to stress tolerance across a diverse collection of strains.

Listeria monocytogenes strains are classified in at least three distinct phylogenetic lineages. There are correlations between lineage classification and source of bacterial isolation; e.g., human clinical and food isolates usually are classified in either lineage I or II. However, human clinical isolates are overrepresented in lineage I, while food isolates are overrepresented in lineage II. sigma(B), a transcriptional regulator previously demonstrated to contribute to environmental stress responses and virulence in L. monocytogenes lineage II strains, was hypothesized to provide differential abilities for L. monocytogenes survival in various niches (e.g., food and human clinical niches). To determine if the contributions of sigma(B) to stress response and virulence differ across diverse L. monocytogenes strains, DeltasigB mutations were created in strains belonging to lineages I, II, IIIA, and IIIB. Paired parent and DeltasigB mutant strains were tested for survival under acid and oxidative stress conditions, Caco-2 cell invasion efficiency, and virulence using the guinea pig listeriosis infection model. Parent and DeltasigB mutant strain transcriptomes were compared using whole-genome expression microarrays. sigma(B) contributed to virulence in each strain. However, while sigma(B) contributed significantly to survival under acid and oxidative stress conditions and Caco-2 cell invasion in lineage I, II, and IIIB strains, the contributions of sigma(B) were not significant for these phenotypes in the lineage IIIA strain. A core set of 63 genes was positively regulated by sigma(B) in all four strains; different total numbers of genes were positively regulated by sigma(B) in the strains. Our results suggest that sigma(B) universally contributes to L. monocytogenes virulence but specific sigma(B)-regulated stress response phenotypes vary among strains.

Transcriptional profiling of the L. monocytogenes PrfA regulon identifies six novel putative PrfA-regulated genes.

The transcriptional activator Positive Regulatory Factor A (PrfA) regulates expression of genes essential for virulence in Listeria monocytogenes. To define the PrfA regulon, the 10403S wildtype (WT) strain, a constitutively active prfA* mutant, and an isogenic ∆prfA mutant were grown under PrfA-inducing conditions in a medium containing glucose-1-phosphate and pre-treated with 0.2% activated charcoal. RNA-seq-generated transcript levels were compared as follows: (i) prfA* and WT; (ii) WT and ∆prfA and (iii) prfA* and ∆prfA. Significantly higher transcript levels in the induced WT or constitutively active PrfA* were identified for 18 genes and 2 ncRNAs in at least one of the three comparisons. These genes included: (i) 10/12 of the genes previously identified as directly PrfA-regulated; (ii) 2 genes previously identified as PrfA-regulated, albeit likely indirectly; and (iii) 6 genes newly identified as PrfA-regulated, including one (LMRG_0 2046) with a σA-dependent promoter and PrfA box located within an upstream open reading frame. LMRG_0 2046, which encodes a putative cyanate permease, is reported to be downregulated by a σB-dependent anti-sense RNA. This newly identified overlap between the σB and PrfA regulons highlights the complexity of regulatory networks important for fine-tuning bacterial gene expression in response to the rapidly changing environmental conditions associated with infection.

Effects of oral administration of aflatoxin B1 and fumonisin B1 in rabbits (Oryctolagus cuniculus).

The effects of prolonged oral administration (21 days) of fumonisin B(1) (FB(1)) and aflatoxin B(1) (AFB(1)) were studied in male New Zealand rabbits by clinical, pathological, biochemical and sphingolipid analyses. Twenty-four animals were randomly divided into the following four experimental groups: (A) 0 mg FB(1)+0 microg AFB(1)/(kg body weight(bw)day) (control); (B) 0 mg FB(1)+30 microg AFB(1)/(kg bw day); (C) 1.5 mg FB(1)/(kg bw day)+30 microg AFB(1)/(kg bw day); (D) 1.5 mg FB(1)/(kg bw day)+0 microg AFB(1). Animals from group B and principally from group C presented clinical signs of intoxication. Rabbits from group C presented a lower body weight gain than controls. Differences were observed between intoxicated rabbits and controls with respect to absolute and relative liver and kidney weight, hepatic function, serum urea and creatinine levels and Sa/So ratio. The most frequent hepatic and renal injuries were vacuolar degeneration of the liver and kidney as shown by the histopathological and serum biochemical results. Combined administration of AFB(1) and FB(1) resulted in synergistic toxic effects both in the liver and in the kidney, but hepatic injuries were more marked.

Effects of seasonal variations and collection methods on the mineral composition of propolis from Apis mellifera Linnaeus Beehives.

The effects of seasonal variations and the methods of collection of propolis produced by Africanized honey bees Apis mellifera Linnaeus, 1758, on the composition of constituent minerals such as magnesium (Mg), zinc (Zn), iron (Fe), sodium (Na), calcium (Ca), copper (Cu), and potassium (K) were evaluated. Propolis was harvested from 25 beehives by scraping or by means of propolis collectors (screen, "intelligent" collector propolis [ICP], lateral opening of the super [LOS], and underlay method). During the one-year study, the propolis produced was harvested each month, ground, homogenized, and stored in a freezer at -10 ºC. Seasonal analyses of the mineral composition were carried out by atomic absorption spectrophotometry and the results were evaluated by analysis of variance (ANOVA), followed by Tukey-Kramer's test to compare the mean values (p<0.05). The results showed that seasonal variations influence the contents of 5 minerals (Mg, Fe, Na, Ca, and Cu), and the propolis harvesting method affects the contents of 4 minerals (Mg, Zn, Fe, and Ca).

Effect of propolis, some isolated compounds and its source plant on antibody production.

Propolis is a beehive product with a very complex chemical composition, widely used in folk medicine because of its several therapeutic activities. Its biological properties and chemical composition may vary according to the geographic location and to the different plant sources. The possible mechanism of action of propolis as well as of its active compounds has been the subject of researchers in recent years. In this work, first we reported the results of our study on the seasonal effect of the immunomodulatory action of propolis on antibody production in bovine serum albumin (BSA)-immunized rats. Then, we compared the effect of Brazilian and Bulgarian propolis, some isolated compounds and Baccharis extract on anti-BSA antibody levels. Based on the results, we conclude that propolis stimulates antibody production, independently of the season and geographic origin. Caffeic acid, quercetin and Baccharis extract had no effect on antibody production, although the importance of isolated compounds is well reported in other biological assays. Propolis action is a consequence of plant-derived products with synergic effects, while isolated compounds or extracts from its plant sources had no effect in this assay.

Comparison of the cost-effectiveness of three approaches to screening for and treating sight-threatening diabetic retinopathy.

The purpose of this study was to analyse and compare the costs involved in screening for and treating sight-threatening diabetic retinopathy in three different clinical settings. In the first setting, diabetologists screened using ophthalmoscopy and color photography, according to the St. Vincent Declaration guidelines, and selected patients for further assessment by a visiting ophthalmologist and for treatment in another hospital. In the second setting, all patients were regularly referred to ophthalmologists, either in the same hospital or elsewhere, for all aspects of eye care. In the third setting, screening was done again with ophthalmoscopy alone by diabetologists who followed the St. Vincent Declaration guidelines; however, further assessment and treatment were carried out in the eye department of the same hospital. Costs to the Italian National Health Service and to patients were calculated per screening performed and per patient subjected to laser treatment as a result of screening. A sensitivity analysis was then performed to simulate the costs of standardised patient populations going through the three different settings. It is concluded that absolute costs would be lower, both for the Italian National Health Service and for patients, if screening, assessment and treatment were all carried out in the same hospital. Equipping a diabetic clinic specially for screening would not be more expensive than delegating eye care to external parties, even for a hospital without an eye department. Moreover, delegating eye care more than doubles costs for patients. Screening for, assessing and treating sight-threatening diabetic retinopathy may be a cost-effective procedure for society as a whole in Italy.

Injection-corrosion cast study of the microcirculation of the brain in haemorrhagic and ischaemic disorders. Preliminary investigation.

The results of studies with the injection-corrosion cast technique in cases of haemorrhagic and ischaemic complications of saccular cerebral aneurysm in man are presented. This technique in conjunction with SEM makes possible a detailed qualitative analysis of the microvascular network. The morphological features of the pathologic areas are analysed. The pathophysiological significance of the observed abnormalities of the cerebral microcirculation are discussed.

Flexor tendon grafts in the fingers.

The authors present the results of twenty-five tendon grafts performed between 1972 and 1982. The end results were evaluated according to these parameters: the pre-operative condition, the donor tendon, the level of the lesion, the age of the patient, and the time elapsed from injury to operation.

[Tunnel syndromes of the upper limb]. / Le sindromi canalicolari dell'arto superiore.

The main canalicular syndromes affecting the upper extremities are reviewed with a description of their aetiopathogenesis. Particular emphasis is placed on the need for early diagnosis and microsurgical treatment that on the basis of personal experience is held to guarantee early recovery.

[Prosthetic treatment of rhizoarthrosis of the thumb]. / Il trattamento protesico della rizoartrosi del pollice.

The aetiopathogenesis of trapeziometacarpal arthrosis or rhizoarthrosis of the thumb is described with particular emphasis on the secondary biomechanical alterations involved. The treatment of the condition and the use of the Swanson technique for the implantation of a Silastic prosthesis are described.

Surgical treatment of Allman type III acromio-clavicular dislocation. A long-term follow-up study.

The authors report the results of a long-term follow-up study of surgical treatment of Allman type III acromio-clavicular dislocation. Despite numerous publications regarding the validity of closed treatment, the authors believe, in accordance with the anatomo functional and pathological knowledge, that open treatment is justified not only in young people but in older ones too. Among the different techniques, the authors prefer acromio-clavicular fixation with threaded wires because the operation time is short and the functional results are good with the lowest number of complications.

[Treatment of subcutaneous rupture of the extensor pollicis longus tendon]. / Sul trattamento delle rotture sottocutanee del tendine estensore lungo del pollice.

The authors present the results of 13 cases operated for subcutaneous rupture of the extensor pollicis longus. Transferral of the extensor indicis proprius was employed in 12 cases and tenodesis involving the extensor pollicis brevis was performed in 1. Results are best when abduction and retropulsion of the thumb are restored. Testing of the correct tension of the transferral procedure is emphasized.

Contributions of σ(B) and PrfA to Listeria monocytogenes salt stress under food relevant conditions.

Listeria monocytogenes is well known to survive and grow under several stress conditions, including salt stress, which is important for growth in certain foods as well as for host infection. To characterize the contributions, to salt stress response, of transcriptional regulators important for stress response and virulence (i.e., σ(B) and PrfA), we analyzed three L. monocytogenes parent strains and isogenic mutants (ΔsigB, ΔprfA, and ΔsigBΔprfA), representing different serotypes and lineages, for their ability to grow, at 25°C, in BHI with 1.9 M NaCl. With regard to growth rate, only the lineage IV strain presented a significant difference between the parent strain and both of its respective mutants lacking prfA (ΔprfA and ΔsigBΔprfA). Conversely, the lineage I and II parent strains showed significantly shorter lag phase in comparison to their respective ΔsigB mutant strains. Intestinal epithelial cell invasion assay and hemolytic activity assays showed a significant role for σ(B) in the former and for PrfA in the latter. To explore the mechanism that may contribute to the extended lag phase in the ΔsigB mutant strain and survival and growth of the parent strain upon salt shock, whole genome transcription profiling was performed to compare transcript levels between the lineage I, serotype 1/2b, parent strain and its isogenic ΔsigB mutant after 30 min of lag phase growth at 25°C in the presence of 1.9M NaCl (salt shock) without aeration. Microarray data showed significantly higher transcript levels for 173 genes in the parent strain as compared to the ΔsigB strain. Overall, 102 of the 173 σ(B) up-regulated genes had been identified in previous studies, indicating that 71 genes were newly identified as being up-regulated by σ(B) in this study. We hypothesize that, among these genes newly identified as σ(B) up-regulated, four genes (lmo2174, lmo0530, lmo0527 and lmo0529) may play a major role in response to salt stress. Lmo2174 contains domains that facilitate sensing and producing a transduction signal in the form of cyclic di-GMP, which may activate the enzymes Lmo0527, Lmo0529 and Lmo0530, which encode proteins similar to those responsible for synthesis of exopolysaccharides that may protect the cell by changing the cell wall structure during salt stress. Overall, our data showed that σ(B), but not PrfA, contributes to growth under salt stress. Moreover, we show that the σ(B) regulon of a L. monocytogenes lineage I strain challenged with salt shock includes salt stress-specific as well as previously unidentified σ(B) up-regulated genes.

σ(B) plays a limited role in the ability of Listeria monocytogenes strain F2365 to survive oxidative and acid stress and in its virulence characteristics.

Listeria monocytogenes strain F2365 was the first strain representative of serotype 4b (lineage I) to be sequenced in 2004, suggesting it could become the model organism for this serotype, which is associated with most human outbreaks of listeriosis worldwide to date. F2365 itself is an outbreak strain that was involved in the listeriosis outbreak associated with Mexican-style soft cheese in California in 1985. In this study, we show through phenotypic and transcriptomic analysis that L. monocytogenes strain F2365 has reduced ability to respond to acid and oxidative stress. F2365 has neither the σ(B)-dependent ability to survive acid or oxidative stress nor the σ(B)-dependent ability to infect Caco-2 epithelial cells in vitro or guinea pigs in vivo. More studies are needed to determine whether the atypical σ(B)-independent response to stress observed in F2365 is strain specific, serotype specific, or even lineage specific.

The effects of Brazilian and Bulgarian propolis in vitro against Salmonella Typhi and their synergism with antibiotics acting on the ribosome.

Salmonella enterica serovar Typhi is the causative agent of typhoid fever in humans, and the use of antibiotics is essential for controlling this infection; however, the excessive use of antibiotics may select resistant strains. Propolis is a honeybee product and its antimicrobial activity has been intensively investigated. Thus, the objective of this study was to investigate a possible synergism between propolis (collected in Brazil and Bulgaria) and antibiotics acting on the ribosome (chloramphenicol, tetracycline and neomycin) against Salmonella Typhi in vitro. The synergism was investigated by using ½ and » of the minimum inhibitory concentration for propolis and these antimicrobial agents, evaluating the number of viable cells according to the incubation time. Brazilian propolis showed a bacteriostatic action against S. Typhi, while Bulgarian propolis showed a bactericidal activity and a synergistic effect with the three antibiotics. Variations in the biological assays might be due to the differences in their chemical compositions. Based on the results, one may conclude that Bulgarian propolis showed an important antibacterial action, as well as a synergistic effect with antibiotics acting on the ribosome, which points out a possible therapeutic strategy evaluating the use of propolis preparations for the treatment of Salmonella Typhi infection.

Antibacterial effects of Brazilian and Bulgarian propolis and synergistic effects with antibiotics acting on the bacterial DNA and folic acid.

Propolis is a honeybee product that has been used since ancient times because of its therapeutic effects. It can be used in the development of alternative therapies for the treatment of many diseases, and because propolis shows antibacterial action, this work was carried out in order to investigate a possible synergism between propolis and antibiotics acting on DNA (ciprofloxacin and norfloxacin) and on the metabolism (cotrimoxazole) against Salmonella typhi. Propolis samples collected in Brazil and Bulgaria were compared in these assays, and the synergism was investigated by using ½ and » of the minimal inhibitory concentration for propolis and antibiotics, evaluating the number of viable cells according to the incubation time. Brazilian and Bulgarian propolis showed antibacterial activity, but no synergistic effects with the three tested antibiotics were seen. Previous works by our laboratory have revealed that propolis has synergistic effects with antibiotics, acting on the bacterial wall and ribosome, but it does not seem to interact with antibiotics acting on DNA or folic acid, and only a bacteriostatic action was seen in these assay conditions.